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Fluorescent mannosides serve as acceptor substrates for glycosyltransferase and sugar-1-phosphate transferase activities in Euglena gracilis membranes
- Ivanova, Irina M., Nepogodiev, Sergey A., Saalbach, Gerhard, O'Neill, Ellis C., Urbaniak, Michael D., Ferguson, Michael A.J., Gurcha, Sudagar S., Besra, Gurdyal S., Field, Robert A.
- Carbohydrate research 2017 v.438 pp. 26-38
- Euglena gracilis, algae, enzymes, fluorescence, high performance liquid chromatography, mass spectrometry, polysaccharides
- Synthetic hexynyl α-D-mannopyranoside and its α-1,6-linked disaccharide counterpart were fluorescently labelled through CuAAC click chemistry with 3-azido-7-hydroxycoumarin. The resulting triazolyl-coumarin adducts, which were amenable to analysis by TLC, HPLC and mass spectrometry, proved to be acceptor substrates for α-1,6-ManT activities in mycobacterial membranes, as well as α- and β-GalT activities in trypanosomal membranes, benchmarking the potential of the fluorescent acceptor approach against earlier radiochemical assays. Following on to explore the glycobiology of the benign protozoan alga Euglena gracilis, α-1,3- and α-1,2-ManT activities were detected in membrane preparations, along with GlcT, Glc-P-T and GlcNAc-P-T activities. These studies serve to demonstrate the potential of readily accessible fluorescent glycans as substrates for exploring carbohydrate active enzymes.